Abstract

The expression of eukaryotic DNA in E. coli will be analyzed employing a recombinant plasmids carrying the qa-2 gene (pVK88) of Neurospora crassa and the his3 (pKK1), his4 (pHis4) and ura3(yIP5) loci of Saccaromyces cervisiae. Employing partial restriction endonuclease digests, the entire qa gene cluster from N. crassa (a group of four contiguous genes) will be cloned into E. coli and analyzed. The 20-200-fold increase in the expression of the catabolic dehydroquinase observed in polynucleotide phosphorylase (pnp) deficient strains of E. coli will be studied both genetically and biochemically. Alterations in the level of transcription, stability of mRNA, and ribosome binding will be examined. The generality of the phenomenon will be tested with recombinant plasmids carrying yeast genes already known to be expressed in E. coli. Attempts will be made to determine if additional N. crassa and S. cervisiae genes can be cloned in high expressing strains which otherwise would not be expressed in E. coli. Similar cloning experiments will be carried out with nuclear DNA from Aspergillus nidulans and Euglena gracilis. Plasmids containing cDNA copies of genes from higher organisms will also be tested for functional expression in mutationally altered E. coli strains.

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM028760-05
Application #
3276046
Study Section
Microbial Physiology and Genetics Subcommittee 2 (MBC)
Project Start
1979-04-01
Project End
1987-11-30
Budget Start
1985-12-01
Budget End
1986-11-30
Support Year
5
Fiscal Year
1986
Total Cost
Indirect Cost

  Institution

Name
University of Georgia
Department
Type
Schools of Arts and Sciences
DUNS #
City
Athens
State
GA
Country
United States
Zip Code
30602

  Publications

Ow, Maria C; Liu, Qi; Mohanty, Bijoy K et al. (2002) RNase E levels in Escherichia coli are controlled by a complex regulatory system that involves transcription of the rne gene from three promoters. Mol Microbiol 43:159-71
Mohanty, B K; Kushner, S R (2000) Polynucleotide phosphorylase, RNase II and RNase E play different roles in the in vivo modulation of polyadenylation in Escherichia coli. Mol Microbiol 36:982-94
Mohanty, B K; Kushner, S R (2000) Polynucleotide phosphorylase functions both as a 3' right-arrow 5' exonuclease and a poly(A) polymerase in Escherichia coli. Proc Natl Acad Sci U S A 97:11966-71
Ow, M C; Liu, Q; Kushner, S R (2000) Analysis of mRNA decay and rRNA processing in Escherichia coli in the absence of RNase E-based degradosome assembly. Mol Microbiol 38:854-66
Mohanty, B K; Kushner, S R (1999) Residual polyadenylation in poly(A) polymerase I (pcnB ) mutants of Escherichia coli does not result from the activity encoded by the f310 gene. Mol Microbiol 34:1109-19
Mohanty, B K; Kushner, S R (1999) Analysis of the function of Escherichia coli poly(A) polymerase I in RNA metabolism. Mol Microbiol 34:1094-108
Zhang, G; Deng, E; Baugh, L et al. (1998) Identification and characterization of Escherichia coli DNA helicase II mutants that exhibit increased unwinding efficiency. J Bacteriol 180:377-87
Wang, R F; O'Hara, E B; Aldea, M et al. (1998) Escherichia coli mrsC is an allele of hflB, encoding a membrane-associated ATPase and protease that is required for mRNA decay. J Bacteriol 180:1929-38
Granger, L L; O'Hara, E B; Wang, R F et al. (1998) The Escherichia coli mrsC gene is required for cell growth and mRNA decay. J Bacteriol 180:1920-8
Zhang, G; Deng, E; Baugh, L R et al. (1997) Conserved motifs II to VI of DNA helicase II from Escherichia coli are all required for biological activity. J Bacteriol 179:7544-50

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